Human practices

Human Practices

Talking with the experts

Before anything else, we would like to mention and also thank our PIs (Mgr. Pavel Dvořák,Ph.D., Ing. RNDr. Martin Marek, Ph.D. and Mgr. Karel Říha, Ph.D.) who fearlessly threw themselves into another year of iGEM competition with us once again, and with whom we discussed the design of our project and workflow during the wet lab experiments.

The design of this year's project presented some challenges that we had to deal with. For this reason, it was necessary to reach out to a larger number of experts on the topics. However, as we were working once more with the same organism ( Bacillus subtilis ) and tried to solve the cyanobacteria problem again, just with a new perspective. We could partly use the contacts from last year. We also needed to see if our project could be translated into practical applications. Lastly, we wanted to get a biological and legal perspective on GMO regulation in the EU, which was our focus in Excellence in another area.

Molecular biology of Bacillus subtilis

Experts: Mgr. Libor Krásný, Ph.D. and RNDr. Mgr. Hana Šanderová, Ph.D.

We chose to work with Bacillus subtilis again, because of our experience and also by meeting our conditions on the organism. After their great help last year, we decided to contact Dr. Krásný and Mrs. Šanderová from the Laboratory of Microbial Genetics and Gene Expression at Czech Academy of Sciences, which is headed by Dr. Krásný, again. They have many years of experience in working with Bacillus subtilis and we were very happy that they were willing to help us again.

With Dr. Krásný and Mrs. Šanderová we firstly consulted the design of our constructs. Especially regarding our inducible repressor in Composite part D (BBa_K3831033), where they recommended the use of the Hyper-spank promoter. In the later stage of wet-lab experiments they helped us with troubleshooting our measurements on the FluoroMax Plus. They even sent us positive controls of Bacillus subtilis with GFP integrated into the chromosome or with a plasmid containing GFP.

Besides the above mentioned, it should also be added that it was thanks to them that we were able to obtain the strain of Bacillus subtilis with which we performed our wet-lab experiments.

Situation regarding cyanobacteria

Experts: Mgr. Rodan Geriš

Regarding the cyanobacteria overpopulation problem we already knew who to contact. And that person is Dr. Geriš, who works in Povodí Moravy, s.p., which manages all watercourses in Moravia region.

Dr. Rodan is an expert on water analysis with a focus on algae and cyanobacteria. First of all, he assured us that cyanobacterial blooms are a constant problem not only in the Czech Republic. Apart from the health risk, we discussed the economic difficulties, due to costly cleaning methods, fish kills, or the loss of tourists. Which is mainly due to the strong smell associated with the decomposition of excessive biomass.

We have learned that in order to prevent the explosive growth of cyanobacteria, we would have to reduce the concentration of phosphorus in the waters to 20-30 μg/l. This is a threshold above which the growth rate no longer changes enough, so reducing it below this level is a critical point with the method of fighting cyanobacteria by removing phosphorus from the waters.

Lastly, we discussed the most appropriate location for our modified organism. We were drawn to stagnant waters where phosphorus sedimentation occurs. Water treatment plants offer the most suitable location for phosphorus uptake, but it is important to consider the adsorption of heavy metals onto the cell wall of Bacillus subtilis. Either this problem would have to be circumvented somehow or application into rivers can be considered.

Applicability of our project

Experts: Doc. Ing. Pavel Ryant, Ph.D. and Ing. Miroslav Florián, Ph.D.

An important goal of our project is the subsequent applicability of the accumulated phosphorus in Bacillus subtilis. For this reason we decided to contact Dr. Ryant, who is the Dean of the Faculty of Agronomy at Mendel University in Brno, together with Dr. Florian from the Central Institute for Supervising and Testing in Agriculture.

Soils in the Czech Republic and elsewhere are gradually running out of phosphorus available for crops. Which would not be such a problem if the total supply of phosphorus for fertilisation were not decreasing along with it. Therefore, any methods for obtaining phosphorus from different sources are more than welcome. Moreover, to our advantage, we have found that there is no biological system in the Czech Republic that accumulates phosphorus for its subsequent reuse. Our system would certainly not be able to accumulate a quantity of phosphorus equal to chemical fertilisers. However, this is not at all a problem for two reasons: we are not trying to create competition with chemical fertilisers, but to come up with something that is partly self-sustainable. And any amount of phosphorus taken out of the water which is transferred back to the field is useful. In addition, our product could be used in organic farming as opposed to chemical fertilizers.

As for the necessary treatment of Bacillus subtilis, we found that heat treatment would be the most suitable and also the easiest. In addition, the unnecessary treatment of the phosphorus form that would be accumulated in our BMCs also plays in our favour, as the soil microflora can cope with this over time. This would reduce the subsequent cost of our product to a minimum. Of course, it is possible to treat the thermally inactivated biomass with acids that would cause the formation of a more active, more readily acceptable, form of phosphorus. However, due to the higher costs, this is not our goal.

The most gratifying circumstance was the desire on the part of Dr. Ryant, if successful, for further cooperation in the form of testing the effectiveness of our fertilizer.

GMO regulation in European Union (EU)

Experts: Mgr. Karel Říha, Ph.D. and Mgr. Et Mgr. Michal Bobek, Diploma, M.Jur., MRes., Ph.D.

For Excellence in another area, we decided to show the flaws in GMO regulation in the EU. In addition to bringing the directives closer, we also wanted an expert opinion on the matter, from the perspective of science and law. The law perspective was given by Dr. Bobek who has served as the Advocate General at the Court of Justice of the EU (CJEU) between 2015-2021. And more regarding this is said on the Excellence in another area page. The science perspective was provided by Dr. Říha. In addition to being one of the PIs for our project, he is part of the Think-tank that is trying to convince the EU not to apply the same restrictive laws to the CRISPR-Cas9 method as to other genome editing methods. In addition, he is the Deputy Director for Research of the Ceitec institute, which has just been highlighted as a good example of how to intervene against the CRISPR-Cas9 restrictions.

Looking firstly at the potential problems, the discussion suggested that it would be preferable to examine genetically modified (GM) products on a case-by-case basis rather than by the methods used, as is the case in other non-EU countries, for example. In the first instance, there is a risk of an outflow of know-how on genome editing outside the EU, as it will be difficult to find funding for projects trying to transfer editing from model organisms to the crops used. Furthermore, there is a risk of economic disadvantage for the EU as imported CRISPR-Cas9 edited crop products that undergo multiple editing will be indistinguishable from unedited crop products. This will put EU farmers at a disadvantage, as adapted crops have higher yields and can grow in more inhospitable areas.

A further often voiced reason for the introduction of CRISPR-Cas9 restrictions is the fear of a monopoly on adapted crops, but this is refuted by, for example, Argentina. Where, by dealing with GM products on a case-by-case basis, they have achieved greater collaboration between farmers and emerging companies in universities.

Lastly, Dr. Říha mentioned that it is very important to spread more awareness amongst genome editing methods to avoid spreading fear of the unknown through public. And this is also one of the reasons why we created our lecture GMO Under the Magnifying Glass.

Talking to the general public

Our team prides itself on two-way communication with the general public. As part of our educational activities, we provide others with source-based information, on the basis of which we then build a discussion on the general possibilities, risks and benefits of GMOs. However, we are also interested in the public's opinion on specific issues related to our project. We have therefore created two public opinion surveys, the results of which have been taken into account at several levels of our project (Integrated Human Practices).

Integrated Human Practices: When creating the project for 2021, we took into account some negative responses to the fact that our CYANOTRAP project (2020) was addressing only the effect of the problem (= cyanobacterial overgrowth) and not its cause (= high phosphate concentrations in surface waters that allow cyanobacteria to overgrow). That is why we decided to focus this year's project on the phosphorus cycle in nature.

Public opinion poll - March 2021

In March, our team was trying to find the theme for this year's project. We wanted to address an important issue affecting many people, so that society would perceive our project as beneficial and meaningful. We asked a wide range of questions to get the public's perspective on many aspects of our work.

Integrated Human Practices: Thanks to the results of the survey, we not only decided on the topic of the project, but also came up with an activity on Excellence in another area that would benefit both us and the public. We were also inspired by the results of the survey to add some content to the lecture GMO Under the Magnifying Glass. In it, we focused on presenting enough verified and substantiated information on issues on which a large number of respondents do not have a strong opinion. Thus, we have prepared quality materials on the basis of which the public will be able to form their own opinion in an informed way. In the following paragraphs we describe what we learned from each question and how it influenced our project.

Introduction of the questionnaire

At the beginning, the respondents were briefly introduced to our team and the purpose of the questionnaire by the following text:

This quiz is designed to survey public opinion on various aspects of our project.

We are a team of students from Masaryk University, Brno University of Technology and Comenius University determined to successfully represent the Czech Republic at iGEM - an international synthetic biology competition!

Last year we managed to win a gold medal and place among the top 5 environmental projects in our age category. In addition to the iGEM competition, we focus on popularizing science using our association Generation Mendel z.s., which is why your opinion is very important to us!

Statistical data

A total of 278 respondents completed our first questionnaire. All of them declared that they were over 15 years old, agreed to processing their responses anonymously for the purposes mentioned above, confirmed that they were free to ask any questions about the project or the survey, and acknowledged that their participation was voluntary and that they could stop completing the survey at any time.
Figure 1: Graphical representation of responses to Question 1 in the Public opinion poll - March 2021.

Question 2: To which gender identity do you most identify?

Of the 278 respondents, 67 identify as a man, 210 as a woman and 1 as other gender. We are aware that it would have been useful to have respondents with a more even and representative gender distribution. We see the reason for the difference in gender representation in the fact that mostly women follow us on social media, where we had shared and promoted the questionnaire.
Figure 2: Graphical representation of responses to Question 2 in the Public opinion poll - March 2021.

Question 3: What is your age?

60.1% of respondents fall into the age category 20-25 years and the other age categories are also unevenly represented. The reason for the difference in the representation of age groups is that we are mainly followed by our friends and classmates on social media, where the questionnaire was shared and promoted.
Figure 3: Graphical representation of responses to Question 3 in the Public opinion poll - March 2021.

Question 4: What is the highest degree or level of school you have completed?

We also asked our respondents about their highest degree or level of education they have completed. The diversity in this parameter is better distributed than in the previous cases, but there is still one prevailing category - 37.1 % of our respondents currently study at a university with a major other than Science. The other categories are more or less evenly distributed.
Figure 4: Graphical representation of responses to Question 4 in the Public opinion poll - March 2021.

Main part of the questionnaire

In the main part of the questionnaire, we asked respondents a total of 14 questions focusing on several smaller topics. We tailored the results from these questions to our project at several levels, which will be described for the critical questions.

Question 5: To which application should we direct our work?

Question 5 asks about the sector of human activity that should benefit from our project. 54% of respondents agreed that we should focus on ecological issues, while 28.8% of respondents believe that we should work on a project whose outcome would be applicable to agriculture. The last two places were occupied by industry (9.4%) and personal use (7.9%).

Integrated Human Practices: The feedback from last year's project solidified our decision to focus on the phosphorus cycle in nature. But how to grasp this problem? We could have developed a system to eliminate phosphate leakage from factories, a system to retain phosphate from fertilizers in fields to prevent it from washing into water bodies, or a system for domestic use that would capture phosphate from washing powder, or water flowing out of the washing machine. However, based on the answers to this question, we opted for an environmentally focused system that would capture phosphates from all three of the above sources from water bodies, thus doing the most to help protect the ecosystem.

As we worked on the project, we also realized that we could connect more of the areas in our project. Our ultimate goal would be to connect them all. We would like to create a device that would capture phosphate from water bodies, the captured phosphate would then be recycled and reused as a fertilizer for agricultural or domestic use, or as a source of phosphorus for industry. For more information, see Implementation.
Figure 5: Graphical representation of responses to Question 5 in the Public opinion poll - March 2021.

In questions 6, 7 and 8 we wanted to check whether public interest in the issue of environmental pollution and in particular cyanobacteria overpopulation is as high as last year.

Question 6: Please rate how strongly you perceive environmental pollution as a factor that harms your health.

From the answers to Question 6, we learned that almost 70% of respondents perceive environmental pollution as a factor with a major impact on their health (values 4 and 5).

Integrated Human Practices: From the results of this question we found that the public is more likely to perceive a link between the state of the environment and their health. This suggests to us that a project addressing cyanobacteria overgrowth (and thus reducing the amount of cyanobacterial toxins in the water) could be perceived by the public as positive and therefore may be easier to take to the implementation stage. With this information, we were reassured that we had chosen an appropriate project topic.

Figure 6: Graphical representation of responses to Question 6 in the Public opinion poll - March 2021.

Question 7: Do you see cyanobacteria overpopulation as a big problem?

Question 8: Do you consider cyanobacteria pollution to be a threat to your health?

Questions 7 and 8 already ask specifically about cyanobacteria. These two similar questions explore the public's view of cyanobacterial overgrowth per se and of cyanobacterial overgrowth as a threat to human health. We find that nearly 80% of respondents perceive cyanobacterial overgrowth as a major problem, and as many as 87.1% perceive cyanobacterial overgrowth to be a threat to their health.

Integrated Human Practices: The responses to these two questions reinforced our belief that following up on last year's project theme was the right choice.

Figure 7: Graphical representation of responses to Question 7 in the Public opinion poll - March 2021.
Figure 8: Graphical representation of responses to Question 8 in the Public opinion poll - March 2021.

Question 9: Have you heard that phosphorus in waters contributes to cyanobacteria overgrowth?

In Question 9, we asked about public awareness of the link between phosphate levels in water and cyanobacteria overgrowth. It showed us that only 56.1% of respondents had ever heard of phosphate in water contributing to cyanobacteria overgrowth.

Integrated Human Practices: The takeaway from this question is that it is important to increase public awareness of this issue. We have written about the link between excessive phosphates in water on our social media and this topic was also mentioned in our GMO Under the Magnifying Glass lecture.

Figure 9: Graphical representation of responses to Question 9 in the Public opinion poll - March 2021.

Question 10: What human activity do you think is most responsible for excess phosphorus in water in the Czech Republic?

In Question 10, we asked how informed the public is about the sources of phosphate in surface waters. According to the Integrated Pollution Register of the Ministry of the Environment of the Czech Republic, the most significant anthropogenic phosphorus emissions include the use of phosphate fertilizers and polyphosphates in detergents [1]. Out of our 278 respondents, 43% think that detergents are involved and 33.6% consider agriculture as the largest source of phosphate pollution. Thus, only 23.4% of respondents have incorrect information about this issue. Even so, we believe that this number needs to be reduced even further.

Integrated Human Practices: We discuss various sources of phosphate pollution with the public both on our social networks and in the GMO Under the Magnifying Glass lecture.

Figure 10: Graphical representation of responses to Question 10 in the Public opinion poll - March 2021.

The rest of the questions were not related to the actual topic of the laboratory part of our project, but were used to find out information we needed for other aspects of our work. They were mainly related to public awareness and opinions on the regulation of GMO use in the European Union and on GMO food.

Question 11: How do you evaluate the regulation of GMO in the European Union?

In Question 11 we asked about the respondents' opinion on the regulation of GMO use in the European Union. 12.6% of respondents think that the regulation of GMOs in the EU is too strict, 15.5% think that it is adequate and 9.7% think that it is insufficient. However, 62.2% of respondents have no opinion on this issue.

Integrated Human Practices: More than a half of the respondents have no opinion on the stringency of regulation of GMO use in the European Union. Since people form their opinion based on the available information, we decided to make it available to people. We have looked at a large number of sources and summarised the relevant information both in the Excellence in another area and in our lecture GMO Under the Magnifying Glass, where we devote a large chapter to this issue. In this way, we want to provide the public with verified and substantiated information in a comprehensible form on the basis of which they can form their own opinion if they wish to do so.

Figure 11: Graphical representation of responses to Question 11 in the Public opinion poll - March 2021.

Question 12: Do you think national governments should become more involved in addressing environmental crises?

Question 12 asks for respondents' views on the involvement of national governments in addressing environmental crises. We find that 97.5% of respondents think that national governments should be more involved in this issue than they have been.
Figure 12: Graphical representation of responses to Question 12 in the Public opinion poll - March 2021.

Question 13: Do you know any specific example of GMO use? If so, what is it?

In the open-ended Question 13, we ask whether respondents know any GMOs. 120 respondents (43.17%) stated that they do not know any example of GMOs. Responses from other respondents varied widely in the degree of accuracy of the description of the GMO example - from the general and misleading "vegetables and fruits" to the precise "Golden Rice, BT corn". The vast majority of respondents mentioned just food GMOs, with only a few cases mentioning laboratory use of GMOs or preparation of insulin.

Integrated Human Practices: In the lecture GMO Under the Magnifying Glass we describe, for example, the preparation of insulin using GMOs and compare it with the previously used isolation of insulin from pig pancreas, and we also talk about the preparation of new supermaterials - for example spider silk in goat milk. This is our way to make people more aware of different uses of GMOs.

Question 14: Do you think GMO foods should be available on the market?

In Question 14 we ask respondents whether they think GM foods should be available on the market. 37.4% of respondents are of the opinion that modified foods should appear on the shelves, 21.2% are against. 41.1% of respondents remain undecided.

Integrated Human Practices: Since people form their opinion based on the information available, we decided to make it available to them. We have searched a large number of sources and summarized important information from them in the lecture GMO Under the Magnifying Glass , where we explain different methods of plant and animal breeding, compare them with genetic engineering methods and above all give objective information on the myths that circulate about GM food. In this way, we want to provide the public with verified and substantiated information in an understandable form, on the basis of which they will be able to form their own opinion if they wish to do so.

Figure 13: Graphical representation of responses to Question 14 in the Public opinion poll - March 2021.

Question 15: Do you think that foods containing GMO should be specially labelled?

Question 15 follows on from the previous question. Assuming that GM foods would be allowed to be sold, respondents are asked whether these foods should be specially labelled. Over 85% of respondents think that GM foods should carry a special label that would easily distinguish them from conventional foods, 14.7% think that this distinction is not necessary.
Figure 14: Graphical representation of responses to Question 15 in the Public opinion poll - March 2021.

Question 16: Would you buy GM food or other GMO products?

Question 16 builds on the previous questions. Here we were interested in whether the respondents would buy GM food or another GMO product. We found that 55% of respondents would buy a GMO product, 45% would not.
Figure 15: Graphical representation of responses to Question 16 in the Public opinion poll - March 2021.

Question 17: What is your opinion on laboratory substitutes for meat and other animal products?

At the time the questionnaire was published, there was a lot of talk about synthetic meat. We were interested in how the public views this particular GMO product, because its production can be beneficial both from an ecological and ethical point of view. We found that 37.1% of respondents had a positive opinion of GM meat substitute products and 29.5% had a negative opinion. 33.5% of respondents have no opinion.

Figure 16: Graphical representation of responses to Question 17 in the Public opinion poll - March 2021.

Question 18: Do you think that a certain genetic modification can significantly change the characteristics of agricultural food? For example, taste or colour?

In the last question of the March questionnaire, we asked the public if they think genetic modification can significantly change the characteristics of food - for example, taste or colour. We wanted to know how the public actually perceives genetic modification and whether they perceive its possibilities and whether it has any limits. We found that 86.3% of respondents think it is possible to fundamentally change the properties of food through genetic modification, 13.7% think it is not possible in the real world.

Integrated Human Practices: In our lecture GMO Under the Magnifying Glass we included a description of the preparation of genetically modified organisms and the differences between GMO and organisms created by classical breeding to make it easier for the public to imagine these processes and better understand their possibilities and limits.

Figure 17: Graphical representation of responses to Question 18 in the Public opinion poll - March 2021.

Public opinion poll - September 2021

By September, we had already done most of the lab work and had a clear idea of how our project should work on a biological level. However, we had several possible visions for how our system could be used in the real world. So we decided to reach out to the public again and see which of our ideas would be best received and most likely to make it to market.